Currently, there are no replacement tooth therapies that provide the function and sensory responsiveness of natural teeth. Our long-term goal is to design and develop functional bioengineered, vital, whole tooth constructs for clinical applications as an improved alternative to synthetic dental implants. The objective of the proposed study is to fabricate biomimetic tooth constructs of specified size and shape using post-natal dental cells encapsulated in gelatin methacrylamide (GelMA) hydrogel scaffolds. We hypothesize that ?tunable? GelMA hydrogel constructs will promote improved dental cell interactions leading to the formation of mineralized dental tissues. The rationale for the proposed research includes the significant health need, our strong preliminary data, and the need for alternatives to currently used replacement tooth therapies. Results from the proposed studies could also be used to develop other bioengineered tissue and organ systems that require proper epithelial and mesenchymal cell interactions. We will test the hypothesis with the following specific aims: 1. Identify growth factors that can be incorporated into GelMA constructs to promote dental cell differentiation and mineralized dental tissue formation. 2. Identify scaffold design for improved DE cell differentiation and enamel production. 3. Characterize biomimetic GelMA tooth bud constructs grown in an in vivo rat jaw implant model. The analyses that we will use to investigate the effectiveness of these different fabrication strategies of GelMA tooth bud constructs include 3D radiographic techniques, and histological and immunohistochemical methods, to measure the extent of polarized dental cell morphology, differentiation, and organized construct mineralization. The expected outcomes of the proposed study include an improved biomimetic 3D tooth bud model that facilitates the formation of organized, functional, bioengineered teeth of specified size and shape. Validation of a functional biomimetic 3D tooth bud will have a significant impact on the tooth tissue engineering field by providing a clinically relevant alternative to artificial dental implants. The contribution of the proposed research is significant as it aims to enhance the quality of dental and oral health for people suffering from tooth loss as well as contribute to other studies aiming to bioengineer similar organs and tissues.